Derivation of ES-like cell from neonatal mouse testis cells in autologous sertoli cells co-culture system

ABSTRACT

Spermatogonial stem cell [SSC] is a self-renewing population of male adult stem cell. SSCs have a differentiation potential which are similar to embryonic stem cells. These Embryonic stem like [ES-like] cells can be a potential source for pluripotent cells for stem cell-based therapy. This study presents an economical and simple co-culture system for pluripotent stem cells generation from neonatal mouse testis. Isolated testicular cells were cultured in DMEM/F12. Characteristics of the isolated cells and obtained ES-like cell were immune-cytochemically confirmed by examining the presence of PLZF, vimentin, Oct4 and Nanog protein. Expression of the pluripotency and germ-cell specific genes was analyzed by qPCR in derived ES-like colony and SSCs respectively. The experiment results indicated that our method of obtaining pluripotent ES-like cells from spermatogonial cells [SCs] is simpler than the described methods. ES-like cells were immunopositive for pluripotency markers. ES-like cell qPCR results indicated significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. The results indicated that ES-like cell with pluripotency characteristic were generated from freshly isolated spermatogonial cells. The pluripotent stem cells provide a cellular reservoir usable for regenerative medicine instead of embryonic stem cells.